Development of an Enzyme-Linked Immunosorbent Assay Method To Detect Mustard Protein in Mustard Seed Oil

2007 ◽  
Vol 70 (1) ◽  
pp. 179-183 ◽  
Author(s):  
STEF J. KOPPELMAN ◽  
RIEK VLOOSWIJK ◽  
GINA BOTTGER ◽  
GERT van DUIJN ◽  
PETER van der SCHAFT ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Seed Oil ◽  
Food Product ◽  
Cross Reactivity ◽  
Polyclonal Antiserum ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mustard Seed ◽  
Calculated Level ◽  
Mustard Protein

An enzyme-linked immunosorbent assay for the detection of mustard protein was developed. The assay is based on a polyclonal antiserum directed against a mixture of mustard proteins raised in rabbits. The assay has a detection limit of 1.5 ppm (milligrams per kilogram) and is suitable for the detection of traces of mustard protein in mustard seed–derived flavoring ingredients. Limited cross-reactivity testing showed that no other plant proteins reacted significantly. From the animal proteins tested, only milk showed some cross-reactivity. With this sensitive assay, it was shown that refined mustard seed oil produced by steam distillation does not contain detectable amounts of mustard protein. Mustard seed oil is used as a flavoring in very low quantities, typically between 40 and 200 mg/kg. Thus, 100 g of a food product flavored with 200 mg of mustard seed oil per kg containing <1.5 mg of protein per kg would represent an amount of mustard seed protein of <30 ng. Taking into account the published literature on allergic reactions to the unintended ingestion of mustard, this conservatively low calculated level indicates that it is unlikely that food products containing mustard seed oil as a flavoring ingredient will elicit an allergic reaction in mustard-allergic individuals.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay Method for the Detection of Rhein in Rheum officinale

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Min Chen ◽  
Tie-Gui Nan ◽  
Jie Xin ◽  
Li Cui ◽  
Bo Zhang ◽  
...  
Keyword(s):  
Quality Control ◽  
Immunosorbent Assay ◽  
High Specificity ◽  
Cross Reactivity ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Simple Method ◽  
Specificity And Sensitivity ◽  
Important Quality ◽  
Aloe Emodin

Rhein is an important quality-control marker of Rheum officinale. The aim of this study was to develop an indirect competitive enzyme-linked immunosorbent assay (icELISA) for rhein detection, which acts as a powerful tool for quality control and proper usage of Rheum officinale. First, a specific and sensitive monoclonal antibody (mAb) against rhein was produced from a stable hybridoma cell line, 1F8, generated by the fusion of mouse myeloma sp2/0 with spleen cells obtained from a Bal b/c mouse immunized with rhein-BSA. Then, an icELISA method was developed with an IC50 value and working range of 0.05 μg L−1 and 0.02–0.11 μg L−1, respectively. The icELISA revealed high assay specificity, since it only had a relatively high cross reactivity with aloe-emodin (27%) and almost no cross reactivity with any other anthraquinones (<1%). When spiked with 0.2–2 mg kg−1 of rhein, the recoveries ranged from 84.19% to 102.90%. Finally, icELISA was used to detect rhein contents of Rheum officinale collected from different regions, and the results corresponded well with those of HPLC. Overall, the developed icELISA with high specificity and sensitivity provided a rapid and simple method for rhein detection, and it may be a powerful tool for quality control and proper usage of Rheum officinale.

scholarly journals Aspergillus Galactomannan Enzyme-Linked Immunosorbent Assay Cross-Reactivity Caused by Invasive Geotrichum capitatum

2006 ◽  
Vol 44 (9) ◽  
pp. 3432-3434 ◽  
Author(s):  
M. Giacchino ◽  
N. Chiapello ◽  
S. Bezzio ◽  
F. Fagioli ◽  
P. Saracco ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Geotrichum Capitatum

Enzyme-linked immunosorbent assay of autoantibodies reacting with thyroid plasma membrane antigens in sera of patients with autoimmune thyroid diseases

1986 ◽  
Vol 113 (2) ◽  
pp. 255-260 ◽  
Author(s):  
Andrzej Gardas ◽  
Kathleen L. Rives
Keyword(s):  
Plasma Membrane ◽  
Immunosorbent Assay ◽  
Lupus Erythematosus ◽  
Antithyroid Drug ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Autoimmune Thyroid Diseases ◽  
Nodular Goitre ◽  
Membrane Antigens ◽  
Toxic Nodular Goitre

Abstract. A sensitive and specific enzyme-linked immunosorbent assay (ELISA) for the detection of autoantibodies reacting with thyroid plasma membrane antigens has been established. Autoantibodies reacting with thyroid plasma membrane antigens were detected by the ELISA in 95% of untreated hyperthyroid Graves', 68% of antithyroid drug-treated Graves' up to four months of the therapy, in 62% of Hashimoto's thyroiditis and in 8.9% of toxic nodular goitre. The ELISA was negative in 100% healthy blood donors, 100% non-toxic nodular goitre, in 12 patients with rheumatoid arthritis, 18 patients with scleroderma and 94% of patients with systemic lupus erythematosus. The mean value of autoantibodies titre was higher in untreated hyperthyroid Graves' (1:84 000) and lowest in positive patients with autoimmune disease of non-thyroid origin (1:4000). The cross-reactivity of antimicrosomal antigen antibodies was below 10%; there was no influence of antithyroglobulin antibodies on the ELISA; and most of the autoantibodies react with plasma membrane antigens different from the TSH binding sites.

Immunological discrimination between a sap-staining fungus and a biological control fungus

1990 ◽  
Vol 68 (7) ◽  
pp. 1578-1588 ◽  
Author(s):  
Brian T. Luck ◽  
Colette Breuil ◽  
David L. Brown
Keyword(s):  
Electron Microscopy ◽  
Biological Control ◽  
Immunosorbent Assay ◽  
Liquid Culture ◽  
Biological Control Agent ◽  
Dry Weight ◽  
Cross Reactivity ◽  
Immunogold Labeling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polyclonal Serum

An enzyme-linked immunosorbent assay (ELISA) was used to detect a sap-staining fungus, Ophiostoma piceae, and a biological-control agent, Gliocladium roseum, grown in liquid culture and in wood. A polyclonal serum prepared against whole cell fragments from broken mycelia of O. piceae detected O. piceae in liquid culture at 0.25 μg dry weight/mL; however, there was moderate cross-reactivity with G. roseum. Antiserum adsorbed on G. roseum had almost no reactivity with G. roseum but still reacted strongly with O. piceae. The specificity of these sera was verified, and the antigenic sites were localized, by immunogold labeling and electron microscopy. These studies confirmed that the adsorbed serum could differentiate between G. roseum and O. piceae and showed that the cell wall was the most reactive cellular component. These results are discussed in relation to the development of immunological probes for the detection of sap-staining and biological control fungi. Key words: polyclonal serum, enzyme-linked immunosorbent assay, immunogold labeling, sap-staining and biological control fungi, electron microscopy.

Inability of Pediococcus pentosaceus to Inhibit Clostridium botulinum in sous vide Beef With Gravy at 4 and 10°C

1994 ◽  
Vol 57 (2) ◽  
pp. 104-107 ◽  
Author(s):  
ALLISON D. CRANDALL ◽  
KAREN WINKOWSKI ◽  
THOMAS J. MONTVILLE
Keyword(s):  
Immunosorbent Assay ◽  
Clostridium Botulinum ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Processed Meat ◽  
Minimally Processed ◽  
False Negatives ◽  
Pediococcus Pentosaceus ◽  
Heat Treated ◽  
Sous Vide

The ability of Pediococcus pentosaceus to inhibit Clostridium botulinum toxigenesis in minimally heat-treated, vacuum-packaged sous vide-type beef with gravy was investigated. The bacteriocinogenic strain P. pentosaceus ATCC 43200 and the nonbacteriocinogenic strain P. pentosaceus 43NP1 were coinoculated with proteolytic and nonproteolytic C. botulinum types A and B spores into minimally processed meat with gravy. Toxin was present in samples inoculated with C. botulinum alone by day 31 at 4°C and by day 6 at 10°C. When coinoculated with C. botulinum, neither strain of Pediococcus was capable of significantly delaying the appearance of toxin. Although an enzyme-linked immunosorbent assay method for botulinal toxin was useful for screening toxin-positive samples, a high proportion of false negatives was revealed by confirmatory mouse bioassays. This research confirms that, if botulinal spores are present, sous vide beef does present a botulinal hazard, even when kept under adequate refrigeration.

scholarly journals Development of a Smartphone-Based Nanozyme-Linked Immunosorbent Assay for Quantitative Detection of SARS-CoV-2 Nucleocapsid Phosphoprotein in Blood

2021 ◽  
Vol 12 ◽  
Author(s):  
Bochao Liu ◽  
Ze Wu ◽  
Chaolan Liang ◽  
Jinhui Lu ◽  
Jinfeng Li ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Cross Reactivity ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Primary Screening ◽  
Global Pandemic ◽  
Novel Coronavirus ◽  
Acid Test ◽  
Control Samples

Since December 2019, a novel coronavirus (SARS-CoV-2) has resulted in a global pandemic of coronavirus disease (COVID-19). Although viral nucleic acid test (NAT) has been applied predominantly to detect SARS-CoV-2 RNA for confirmation diagnosis of COVID-19, an urgent need for alternative, rapid, and sensitive immunoassays is required for primary screening of virus. In this study, we developed a smartphone-based nanozyme-linked immunosorbent assay (SP-NLISA) for detecting the specific nucleocapsid phosphoprotein (NP) of SARS-CoV-2 in 37 serum samples from 20 COVID-19 patients who were diagnosed by NAT previously. By using SP-NLISA, 28/37 (75.7%) serum samples were detected for NP antigens and no cross-reactivity with blood donors’ control samples collected from different areas of China. In a control assay using the conventional enzyme-linked immunosorbent assay (ELISA), only 7/37 (18.91%) serum samples were detected for NP antigens and no cross-reactivity with control samples. SP-NLISA could be used for rapid detection of SARS-CoV-2 NP antigen in primary screening of SARS-CoV-2 infected individuals.

scholarly journals Development of a Sandwich Enzyme-Linked Immunosorbent Assay for Detection and Quantification of Clam Residues in Food Products

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Stef J. Koppelman ◽  
Ashley L. Lardizabal ◽  
Lynn Niemann ◽  
Joe L. Baumert ◽  
Steve L. Taylor
Keyword(s):  
Sandwich Elisa ◽  
Food Product ◽  
Food Products ◽  
Elisa Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Allergic Reactions ◽  
Arctica Islandica ◽  
Limit Of Quantification ◽  
Detection And Quantification

Seafood is a frequent cause of allergic reactions to food globally. The presence of undeclared trace amounts of clam can cause allergic reactions in sensitive individuals. Limited tools are available to test food products for the presence of traces of clam. We report on the development of a sandwich ELISA that can detect and quantify clam protein in food. Antisera against a mix of two commercially important clam species, Atlantic Surf (Spisula solidissima) and ocean quahog (Arctica islandica), were raised in rabbit and sheep. A sandwich ELISA was constructed with this antisera, and sensitivity and specificity were evaluated. Also, model food products spiked with clam protein were analyzed to assess the performance of the ELISA. Comparison was made with a commercially available ELISA for crustacea. The lower limit of quantification of the sandwich ELISA is 2.5 ppm clam protein in food samples, allowing the detection of low amounts of clam that may trigger a reaction in clam allergic patients. The sandwich ELISA was highly specific with cross-reactivity only noted for other molluscan shellfish (mussel and scallop). Clam protein in tomato juice and potato cream soup was detected well with recoveries ranging from 65 to 74% and from 74 to 113%, respectively. However when potato cream soup was retorted, the recover fell to 20%, imposing the risk of underestimating the clam content of a food product. A commercially available crustacean ELISA test was not suitable to detect clam protein. The sandwich ELISA described here is suitable for detection and quantification of clam protein in food products. Care should be taken with food products that have been retorted as the results may be underestimated.

scholarly journals Fecal Cortisol and Progesterone Concentrations in Post Partus of Etawah Crossbreed Goat

2020 ◽  
Vol 151 ◽  
pp. 01031
Author(s):  
Claude M. Airin ◽  
Amelia Hana ◽  
Sarmin Sarmin ◽  
Pudji Astuti
Keyword(s):  
Immunosorbent Assay ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Stress Hormone ◽  
Fecal Samples ◽  
Fecal Cortisol ◽  
Fecal Extract ◽  
Last Stage ◽  
Preliminary Study ◽  
Freeze Dryer

Progesterone (P4) is a dominant hormone during pregnancy. In the later stage of pregnancy, the stress hormone particularly cortisol (C) may increase for initiating the parturition process as a consequence of fetal stress. This study was a preliminary study to compare the concentration of P4 and C in feces of Etawah Crossbreed Goat during their last stage of pregnancy and post partus. This study used 5 pregnant Etawah Crossbreed Goats (t 20th weeks) of pregnancy. Fecal samples were collected in the 20th week of pregnancy to 2 weeks of postpartum. All fecal samples were then dried using a freeze dryer (Labfreez FD10-MR) for 7 days at -80°C. Afterward, dried feces were pulverized and extracted by using 3ml of methanol 80%. The fecal extract was then analyzed the P4 and C concentrations using the enzyme-linked immunosorbent assay method. Concentrations of P4 and C metabolites in the last stage of pregnancy were 5,506.18 3,396.72 ng/g dry feces and 136,625.83 42,479.22 ng/g feces, respectively. Concentrations of P4 and C metabolites in the 2 weeks postpartum decreased at 669.38 P 643.9 ng/g feces and 110,295 / 14,378, 8 ng/g feces, respectively. It canbe concluded that there was a difference in the fecal progesterone and cortisol concentrations between the last phase of pregnancy and the postpartum phase.

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